title: Split intein facilitated tag affinity purification for recombinant proteins with controllable tag removal by inducible auto-cleavage.
abstract: Purification tags are robust tools that can be used to purify a variety of target proteins. However, tag removal remains an expensive and significant issue that must be resolved. Based on the affinity and the trans-splicing activity between the two domains of Ssp DnaB split-intein, a novel approach for tag affinity purification of recombinant proteins with controllable tag removal by inducible auto-cleavage has been developed. This system provides a new affinity method and avoids premature splicing of the intein fused proteins expressed in host cells. The affinity matrix can be reused. In addition, this method is compatible with his-tag affinity purification technique. Our methods provide the insights for establishing a novel recombinant protein preparation system.
authors.affiliation: Institute of Molecular & Experimental Therapeutics, East China Normal University, Shanghai, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Lu
authors.forename: Ziyong
authors.initials: Z
authors.lastname: Sun
authors.forename: Yanchun
authors.initials: Y
authors.lastname: Tang
authors.forename: Junyong
authors.initials: J
authors.lastname: Chen
authors.forename: Fengyuan
authors.initials: F
authors.lastname: Tang
authors.forename: Jing
authors.initials: J
authors.lastname: Zhang
authors.forename: Jian-Ning
authors.initials: JN
authors.lastname: Liu
mesh_terms: D000345:Affinity Labels / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D002846:Chromatography, Affinity / Q000379:methods*; D053488:DnaB Helicases / Q000737:chemistry / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D055767:Immobilized Proteins / Q000737:chemistry / Q000378:metabolism; D047668:Inteins; D013058:Mass Spectrometry; D015202:Protein Engineering / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D013696:Temperature
chemical_list: D000345:Affinity Labels; D055767:Immobilized Proteins; D011993:Recombinant Fusion Proteins; D053488:DnaB Helicases